Biochemistry Lab
Module 5: Polysaccharides
INTRODUCTION
➢ Carbo → carbon; hydro → water
➢ CH2O
➢ Monosaccharide → glucose → most common
o Dextrose → D-glucose → if no appetite to
eat ang patient
o For energy ng body
➢ Bakit hindi nalang puro glucose or glucose directly
yung kinakain ng mga tao?
o Magkakaron kasi ng sugar rush → this lasts
only for a short amount of time → usually
causes energy deficit
o Like kumain ka ng sugar → kung kelan ka
kumain, don lang siya magtatake effect and
wala na reserves for later = energy deficit
5.1 WHAT IS A POLYSACCHARIDE
➢ Long-chain carbohydrate made up of smaller
carbohydrates called monosaccharides
➢ Contains many monosaccharide units bonded to each
other by glycosidic linkages → chains of
monosaccharides
➢ Each monosaccharide is connected together via
glycosidic bonds to form the polysaccharide
➢ 200 to 250 monosaccharides long → linear or
branched carbon chains
➢ Usual structure of polysaccharides → six-carbon
repeating monosaccharides linked together by oxygen
➢ CHEMICAL FORMULA → (C6H10O5)n
o n is a number larger than 40
➢ Polymers → plastics
➢ Not sweet
➢ Do not give a positive results in Tollens, Benedict’s,
and Fehling’s
5.2 TYPES OF POLYSACCHARIDES
CELLULOSE
➢ Biodegradable, odorless, and has no taste
➢ Straight chain polymer → linear polymer
➢ Organic compound
➢ Most abundant polysaccharide
➢ Made up of a linear chain of multiple glucose residues
(300 to 1000 or more units)
➢ Linked by ß(1→4) glycosidic bonds
➢ Microfibril → renders tensile strength to the cell →
serves as the plant’s “cytoskeleton”
➢ Cellulose rich structures → paper → mas durable
BSN 1 | 1st Term
S.Y. 2022 - 2023
by Guia Isabel S. Eugenio
STARCH
➢ Chain of glucose molecules → linear form → α-
glucose
➢ Plants → common storage starch → amylose
➢ Animals → common storage starch → glycogen
o Found in animal liver and muscle
➢ Starch in both plants and animals → used as “fuel”
→ broken down into smaller molecules for energy
production
➢ Starch → linear form → easiest to break down into
energy
➢ Examples are wheats and potatoes
➢ Has two different types:
o Amylose
o Amylopectin
AMYLOSE
➢ Straight-chain glucose polymer (15-20% of starch)
➢ Connected via α(1→4) glycosidic linkage
AMYLOPECTIN
➢ Branched-chain glucose polymer (80-85% of starch)
➢ Either connected via α(1→4) glycosidic linkage or via
α(1→6) glycosidic linkage
GLYCOGEN
➢ One of the main forms of energy reserves
➢ It is a complex carbohydrate that stores excess
glucose
➢ A.k.a. animal starch
➢ It’s called animal starch → amylopectin constituent of
plant starch → similar → composition and structure
→ polysaccharide constituent of glycogen
o Difference → extensive branching in
glycogen at every 8 to 12 glucose
➢ Linked by α(1→6) glycosidic bonds
➢ α-glycosidic bonds form open helical polymers
o ß-glycosidic bonds → produce straight
strands forming structural fibrils
➢ Under the microscope → glycogen → asterisk or star
appearance
➢ Occurs as a granule in the cytosol of the cell
➢ Diameter: 10 to 10 nm2
➢ Core of a glycogen granule → glycogenin → enzyme
that catalyzes the conversion of glucose to glycogen
→ serves as a primer
CHITIN
➢ Similar to cellulose
➢ Linear polymer → no branching
➢ Serves as a protective covering and mechanical
support to soft-bodied organisms producing it
Module 5: Polysaccharides
➢ Insects and arthropods → chitin is a crucial
component for their exoskeleton
➢ Also present → insect body wall, gut lining, salivary
glands, mouth parts, and muscle attachment points
➢ Chitinous fungi → chitin is present in their cell wall
rather than cellulose
➢ Arachnids → chitin is present in their hard outer
covering
➢ Also present in:
o Mollusks → sa radula nila
o Cephalopods → sa beaks nila
➢ Enzymes that degrade chitin → used as potential
fungicides as well as insecticides
➢ Chitin and chitosan → used as:
o Food additive
o Thickener
o Texturizer
o Emulsifier
o Humectant
o Stabilizer in confectionery, beverages, and
other foods
5.3 TESTS FOR POLYSACCHARIDES
FEHLING’S TEST
➢ To test the presence of aldehydes → detected by
reduction of the deep blue solution of copper(II) to a
red precipitate of insoluble copper oxide
➢ NOT specific for aldehydes
➢ Also a test for monosaccharides
➢ Differentiate water-soluble aldehyde and ketone
functional group
➢ No observed changes → no aldehyde
➢ Starch will be hydrolyzed → broken down into
simpler units
➢ Positive siya if may reduction of copper → red →
brick red precipitate (+)
o Aldehydes → reducing agents = reducing
something
o Glucose will reduce copper → gain electron
→ (blue) nabawasan = Cu2+ → Cu+ = red
PROCEDURE
➢ To 2 ml of cooked starch solution in a test tube, add 2
ml of Fehling’s solution and warm over a water bath
for 3-5 minutes
➢ Observe reaction
IODINE TEST
➢ Iodine (iodine-potassium iodide, I2KI) staining
distinguishes starch (poly) from mono, di, and other
polysaccharides
➢ Test for the presence of starch
➢ Starch → coiled polymer of glucose
➢ Iodine → interacts with these coiled molecules and
become bluish black
➢ Iodine dissolved in KI → deep blue-black color
➢ Reaction is the result of the formation of polyiodide
chains from the reaction of starch and iodine
➢ The amylose, or straight-chain portion of starch,
forms helices where iodine molecules assemble,
forming a dark blue-black color
➢ I2 (Iodine) + I- (KI) → I3-
➢ Iodine slides into the starch (amylose) coil to give a
blue-black color
➢ Mas madali ipalipot yung straight na chain
➢ Add Iodine -KI reagent to a solution or directly on a
potato or other materials such as bread, crackers, or
flour → a blue-black color results if starch is present
➢ If di present ang amylose → orange/yellow
➢ If present ang amylopectin → no color → ito yung
cellulose or disaccharides such as sucrose ang
present
o Amylopectin → shorter helices → iodine
molecules unable to assemble →
yellow/orange
➢ Starch → broken down → blue-black color not
produced
➢ Completion of hydrolysis → no color change
o Hydrolysis → hydrogen → addition of
hydrogen
▪ Good sources of hydrogen → acid
OBSERVATION
➢ Cooked starch → dark blue color → presence of
starch (+)
➢ Raw starch → little to no color change (-)
REFLECTION
➢ Cooked starch → starch is present → more
production of amylose
➢ Raw starch → starch is not present / very small
amount → amylose and amylopectin
➢ Hydrolysis → amylose or amylopectin → adding ng
acid → heating = glucose → breakdown → negative
kasi di mapapalibutan → isang glucose lang
➢ Heating → amylopectin will turn into amylose or
produce more amylose
➢ Cooking cause usable nutrients to convert into
harder-to-break-up starch, causing a lack of nutrients
PROCEDURE
➢ PART 1
o Place 1 ml of 2% raw starch (stir the mixture
thoroughly as the starch particles tend to
settle at the bottle) in a test tube, and in
another test tube, and in another test tube 1
ml of 2% cooked starch.
o Place 2 drops of iodine in each test tube and
take note of the color change in iodine.
Module 5: Polysaccharides
o Mix the contents of the two test tubes in one
and place in boiling water bath.
o Observe the gradual change in color of
iodine.
➢ PART 2
o Place 1 ml of 2% cooked starch solution in
test tube 1
o 1 ml of 1% glycogen solution in test tube 2
o place a piece of filter paper on a watch
glass.
o Add two drops of iodine to each.
o Observe color reaction of each with iodine.
HYDROLYSIS OF CELLULOSE + FEHLING’S TEST
➢ Yields beta-D-glucose
➢ Positive results
GLYCOGEN
➢ Storage form of carbohydrates in animals
➢ Highly branched → every 10 carbons → (-) iodine
test
➢ Homopolymer of apha-D-glucose
➢ Heating → straight chain → (+) iodine test
➢ Hydrolyze → broken down → (+) Fehling’s (-) iodine
➢ Ethyl alcohol → precipitating agent
➢ Iodine test → reddish brown color

HYDROLYSIS OF POLYSACCHARIDES
➢ Process of converting polysaccharides into simple
monosaccharide components → saccharification
➢ Utilizes water molecules
PROCEDURE
➢ Place 15 ml of cooked starch solution in a beaker and
add 1 ml of concentrated HCl
➢ Boil the mixture continually until it does not form a
purple complex when tested with iodine
➢ Add 1-2 drops of Phenolphthalein and NaOH drop
wise until neutral as indicated by a pale pink endpoint
➢ Test 2 ml of the hydrolyzed starch with 2 ml of
Fehling’s reagent and warm in water bath for 3-5
minutes.
➢ Compare the results obtained from the starch tested
in procedure B. 5.4 GLYCEMIC INDEX
➢ It is a system of assigning a number to carbohydrate-
containing foods according to how much each food
PARCHMENT PAPER increases blood sugar or blood glucose
PROCEDURE ➢ Low = >55
➢ Dip a strip of filter paper into 6M sulfuric acid solution ➢ Medium = 56-69
for exactly 15 minutes ➢ High = 70<
➢ Transfer the strip immediately into a 3M NaOH
solution for neutralization
➢ Wash the filter paper thoroughly in water. Observe 5.5 SUMMARY
and compare the texture of the treated filter paper COMPONENT RESULTS
with the original strip IODINE TEST
➢ Test the treated “parchment paper” with iodin and Starch (+)
compare with untreated paper and starch Glycogen Without heating (-)
With heating (+)
CELLULOSE Cellulose (Filter Untreated (-)
CELLULOSE TEST paper) Treated (-)
➢ Iodine test with cellulose (filter paper) 2% raw starch (+)
o No reaction 2% cooked starch (+)
o No branching sa cellulose FEHLING’S TEST
CELLULOSE SOLUBILITY Starch Hydrolyzed (+)
➢ Schweizer’s reagent [Cu(NH3)4(H2O)2](OH)2 Unhydrolyzed (-)
o Deep blue compound used in purifying Cellulose Treated (+)
cellulose Untreated (-)
➢ Cotton → 90% cellulose → most indigestible